Aging in a dish: Age-dependent changes of neuronal survival, protein oxidation, and creatine kinase BB expression in long-term hippocampal cell culture
Aging in a dish: Age-dependent changes of neuronal survival, protein oxidation, and creatine kinase BB expression in long-term hippocampal cell culture
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DOI：
10.1002/(sici)1097-4547(19991015)58:2
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发表时间：
1999-10-15
影响因子：
4.2
通讯作者：
Butterfield, DA
中科院分区：
文献类型：
--
作者：
Aksenova, MV;Aksenov, MY;Butterfield, DA
不同实验系统的结果表明，氧化损伤的增加在正常衰老和与年龄相关的病理中起作用。在目前的研究中，海马神经元的长期培养作为一个模型系统进行了研究。根据Gompertz定律，在长期培养中神经元的存活率会下降，并且神经元“在培养皿中衰老”与细胞蛋白质氧化损伤的增加有关。氧化蛋白的Western blot分析和原位免疫细胞化学方法（用于分析固定细胞中蛋白质氧化的方法）均证实老化神经元中蛋白质羰基形成增加。在衰老的神经元培养中，肌酸激酶（CK）含量逐渐增加，但每免疫反应蛋白的酶活性下降，表明无活性CK分子的积累。CK含量的增加不是蛋白质普遍升高的结果，因为β -肌动蛋白含量的分析显示了一种时间依赖性的损失，可能反映了细胞过程数量随着年龄的增长而减少。这些发现显示了“培养皿中的衰老”，与衰老与蛋白质氧化增加有关的概念相一致，为研究与氧化应激相关的与年龄相关的神经退行性疾病提供了一个系统。(C) 1999 Wiley-Liss, Inc。
Results from different experimental systems demonstrate that increased oxidative damage plays a role in normal aging and age-associated pathology. In the current study, long-term cultures of hippocampal neurons were examined as a model system. It was established that neuronal survival in long-term culture decreases according to the Gompertz law and that neuronal "aging in the dish" is associated with increased oxidative damage of cell proteins. The increase of protein carbonyl formation in aged neurons was demonstrated both by Western blot analysis for oxidized proteins and by in situ immunocytochemical method, which was developed to analyze protein oxidation in fixed cells. In aging neuronal cultures, a gradual increase in creatine kinase (CK) content but decreased activity of enzyme per immunoreactive protein was found, suggesting the accumulation of inactive CK molecules, The increase in CK content was not a result of generalized protein elevation, since analysis of beta-actin content showed a time-dependent loss, probably reflecting decreased number of cellular processes with aging. These findings, showing "aging in a dish," consistent with the notion that aging is associated with increased protein oxidation, provide a system for study of age-related neurodegenerative disorders associated with oxidative stress. (C) 1999 Wiley-Liss, Inc.